Microvascular endothelial cell heterogeneity in the central nervous system

中枢神经系统微血管内皮细胞异质性

基本信息

批准号：
7177092
负责人：
Joel S Pachter
金额：
$ 19.11万
依托单位：
UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-01-22 至 2008-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7177092
关键词：
Address Affect Area Attention Behavior Biochemistry Biological Blood Blood - brain barrier anatomy Blood Vessels Blood capillaries Brain Capillary Endothelial Cell Cerebral cortex Cerebrum Classification Condition Coupling DNA DNA Microarray Chip DNA Microarray format Data Depth Development Disease Drug Formulations Endothelial Cells Evaluation Exploratory/Developmental Grant Gene Expression Gene Expression Profiling Genes Goals Grant Health Heterogeneity Homeostasis In Situ Inflammatory Investigation Lead Light Location Microarray Analysis Modeling Molecular Molecular Profiling Nature Neuraxis Neurons Parietal Lobe Pattern Peripheral Permeability Physiological Population Process RNA Rate Reporting Reproducibility Research Risk Sampling Source Spinal Cord Staging Support Contracts System Techniques Testing Tissues Transcript Trees United States National Institutes of Health arteriole base capillary design hemodynamics laser capture microdissection neurovascular unit novel tissue culture tool vascular bed venule

项目摘要

DESCRIPTION (provided by applicant): The issue of microvascular endothelial heterogeneity greatly impacts vascular function in both health and disease, and potentially severely complicates the use of tissue culture-based models to study endothelial behavior. Yet, despite this topic having been explored in considerable depth in peripheral vascular beds, surprisingly little attention has yet been focused on the microvascular axis of central nervous system (CNS). Unrelated reports describing particular genes have, nonetheless, provocatively suggested that a diversity in endothelial gene expression may exist at the segmental level (i.e., between arterioles, capillaries and venules) as well as the regional level of the CNS, and approximate or surpass that found in the periphery. Given the preeminence of the microvascular network in forming both the blood-brain barrier and blood- spinal cord barrier, and the significant microvascular involvement in inflammatory, infectious, degenerative and traumatic conditions of the CNS, it is critical that there be a systematic and detailed evaluation of this clinically important issue. It is of further importance that such analysis be performed on microvascular tissue in situ," as endothelial cell gene expression is exquisitely prone to environmental modulation. Accordingly, we propose the following Specific Aims: 1) To validate and optimize the approach of coupling laser capture microdissection (LCM) of brain microvascular endothelial cells, with global gene expression profiling by DMA microarray analysis. This will include demonstrating feasibility, by maximizing the percentage of transcripts detected (P call rate) in capillaries only, and establishing reproducibility, by identifying both technical and biological sources of variance in microarray data; and 2) To use the LCM/microarray approach to compare the global gene expression profiles of endothelial cells from capillaries, venules and arterioles. The R21 format chosen is specifically designed for projects that are exploratory and may involve considerable risk, but lead to the development of novel techniques that could have major impact on a field of research. In this regard, results obtained here will set the stage for evaluating regional gene expression by the neurovascular unit (i.e., endothelial cells and intimately associated neural cells) along the CNS microvascular tree in both health and disease. In turn, these studies will enable formulation of more precise endothelial models to study the molecular basis of physiological and pathophysiological processes of the cerebral microvasculature.

描述（由申请人提供）：微血管内皮异质性问题极大地影响健康和疾病中的血管功能，并且可能使使用基于组织培养的模型来研究内皮行为变得严重复杂化。然而，尽管这个话题已经在外周血管床中进行了相当深入的探索，但令人惊讶的是，人们对中枢神经系统（CNS）微血管轴的关注却很少。然而，描述特定基因的不相关报告令人兴奋地表明，内皮基因表达的多样性可能存在于节段水平（即小动脉、毛细血管和小静脉之间）以及中枢神经系统的区域水平，并且接近或超过发现的结果在外围。鉴于微血管网络在形成血脑屏障和血脊髓屏障方面的突出作用，以及中枢神经系统炎症、感染、退行性和创伤性疾病中微血管的显着参与，因此系统和详细的研究至关重要评估这一临床重要问题。更重要的是，这种分析要在微血管组织上进行原位，因为内皮细胞基因表达非常容易受到环境调节。因此，我们提出以下具体目标：1）验证和优化耦合激光捕获的方法脑微血管内皮细胞的显微解剖（LCM），通过 DMA 微阵列分析进行全局基因表达谱分析，这将包括通过最大限度地提高毛细血管中检测到的转录本的百分比（P 调用率）来证明可行性。仅，并通过识别微阵列数据中的技术和生物来源的差异来确定可重复性；以及2)使用LCM/微阵列方法来比较来自毛细血管、小静脉和小动脉的内皮细胞的整体基因表达谱。专为探索性项目而设计，可能涉及相当大的风险，但会导致新技术的发展，可能对研究领域产生重大影响。在这方面，这里获得的结果将为评估健康和疾病中中枢神经系统微血管树上神经血管单元（即内皮细胞和密切相关的神经细胞）的区域基因表达奠定基础。反过来，这些研究将能够制定更精确的内皮模型，以研究脑微血管系统的生理和病理生理过程的分子基础。